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虫号: 730283
注册: 2009-03-24
专业: 无机非金属基复合材料

[交流] 分享-Surface Plasmon Resonance-无重复已有1人参与

1. INTRODUCTION 3
2. PRINCIPLES AND APPLICATIONS OF SURFACE PLASMON RESONANCE. 4
2.1. Principles 4
2.2. Applications 5
2.2.1. What SPR is good for 5
2.2.2. What SPR is not good for. 7
3. GENERAL PRINCIPLES OF BIACORE EXPERIMENTS 8
3.1. A typical experiment 8
3.2. Preparation of materials and buffers 9
3.3. Monitoring the Dips 9
4. LIGAND 10
4.1. Direct versus indirect immobilisation 10
4.2. Covalent immobilisation 11
4.2.1. A general approach 11
4.2.2. Choice of chemistry 12
4.2.3. Prepare the protein 12
4.2.4. Pre-concentration 12
4.2.5. Amine coupling 14
4.2.6. Regeneration 15
4.2.7. Adjusting the immobilisation conditions. 16
4.3. Non-covalent immobilisation (ligand capture) 16
4.3.1. Using an existing strategy 17
4.3.2. Developing a new strategy 17
4.4. Activity of immobilised ligand 18
4.5. Control surfaces 19
4.6. Re-using sensor chips 19
5. ANALYTE 20
5.1. Purity, activity and concentration 20
- 1 -
5.2. Valency 20
5.3. Refractive index effect and control analytes 21
5.4. Low molecular weight analytes 22
6. QUALITITATIVE ANALYSIS; DO THEY INTERACT? 22
6.1. Positive and negative controls 22
6.2. Qualitative comparisons using a multivalent analyte 23
7. QUANTITATIVE MEASUREMENTS 23
7.1. Affinity 24
7.1.1. Concepts 24
7.1.2. Experimental design 24
7.1.3. Data analysis 26
7.1.4. Controls 27
7.1.5. Non-linear Scatchard Plots 27
7.2. Kinetics 30
7.2.1. Concepts 30
7.2.2. Experimental design 31
7.2.3. Data analysis 33
7.2.4. Controls 34
7.3. Stoichiometry 35
7.4. Thermodynamics 35
7.5. Activation energy 37
8. CONCLUSION 38
9. APPENDIX 38
9.1. Physical basis of SPR 38
9.2. Samples and buffers 39
9.2.1. Guidelines for preparing samples and buffers for use on the BIAcore 39
9.2.2. Standard buffers 40
9.3. Additional information 41
10. REFERENCES 41
11. ACKNOWLEDGEMENTS 42
13. FIGURE 1. ANALYSIS OF DIPS. 44
14. TABLES 45
14.1. Table 1. BIAcore instruments currently available (January 1999) 45
14.2. Table 2. Sensor chips available for use on the BIAcore 46
14.3. Table 3. Covalent coupling chemistry 47
14.4. Table 4. Techniques for ligand capture. 48
14.5. Table 5. Trivial causes of complex binding kinetics 49
14.6. Table 6. Distinguishing some non-trivial causes of complex kinetics 50

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